Generation of uniform liver spheroids from human pluripotent stem cells for imaging-based drug toxicity analysis

Gyunggyu Lee, Hyemin Kim, Ji Young Park, Gyeongmin Kim, Jiyou Han, Seok Chung, Ji Hun Yang, Jang Su Jeon, Dong Hun Woo, Choongseong Han, Sang Kyum Kim, Han Jin Park, Jong Hoon Kim

Research output: Contribution to journalArticlepeer-review

34 Citations (Scopus)

Abstract

Recent advances in pluripotent stem cell technology provide an alternative source of human hepatocytes to overcome the limitations of current toxicity tests. However, this approach requires optimization and standardization before it can be used as a fast and reliable toxicity screening system. Here, we designed and tested microwell culture platforms with various diameters. We found that large quantities of uniformly-sized hepatocyte-like cell (HLC) spheroids (3D-uniHLC-Ss) could be efficiently and reproducibly generated in a short period time from a small number of differentiating human pluripotent stem cells (hPSCs). The hPSC-3D-uniHLC-Ss that were produced in 500-μm diameter microwells consistently exhibited high expressions of hepatic marker genes and had no significant signs of cell death. Importantly, a hepatic master gene hepatocyte nuclear factor 4α (HNF4α) was maintained at high levels, and the epithelial-mesenchymal transition was significantly attenuated in hPSC-3D-uniHLC-Ss. Additionally, when compared with 3D-HLC-Ss that were produced in other 3D platforms, hPSC-3D-uniHLC-Ss showed significantly higher hepatic gene expressions and drug-metabolizing activity of the enzyme, CYP3A4. Imaging-based drug toxicity studies demonstrated that hPSC-3D-uniHLC-Ss exhibited enhanced sensitivity to various hepatotoxicants, compared to HLCs, which were differentiated under 2D conditions. Precise prediction of drug-induced hepatotoxicity is a crucial step in the early phases of drug discovery. Thus, the hPSC-3D-uniHLC-Ss produced using our microwell platform could be used as an imaging-based toxicity screening system to predict drug hepatotoxicity.

Original languageEnglish
Article number120529
JournalBiomaterials
Volume269
DOIs
Publication statusPublished - 2021 Feb

Bibliographical note

Funding Information:
This research was supported by the National Research Foundation of Korea funded by the Ministry of Science and ICT (Bio and Medical Technology Development Program: No. 2018M3A9H1019504 for JHK) and a Korea University Grant (JHK).

Funding Information:
This research was supported by the National Research Foundation of Korea funded by the Ministry of Science and ICT (Bio and Medical Technology Development Program: No. 2018M3A9H1019504 for JHK) and a Korea University Grant (JHK).

Publisher Copyright:
© 2020 Elsevier Ltd

Keywords

  • Hepatocyte
  • Microwell
  • Spheroid
  • Stem cell
  • Toxicity

ASJC Scopus subject areas

  • Bioengineering
  • Ceramics and Composites
  • Biophysics
  • Biomaterials
  • Mechanics of Materials

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